Conjugated nonadecadienoic acid compositions

ABSTRACT

The present invention provides a method of using conjugated nonadecadienoic acid (CNA), one of its derivatives, or a combination thereof to inhibit lipoprotein lipase activity associated with a cell, to control body fat in a human or non-human animal, to inhibit cyclooxygenase activity in a cell, to inhibit platelet aggregation in a human or non-human animal, and to prevent or treat a condition or disease related to platelet aggregation.

CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of U.S. provisionalapplications bearing Serial No. 60/294,908, filed on May 31, 2001, andSerial No. 60/362,248, filed on Mar. 6, 2002.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] To be determined.

BACKGROUND OF THE INVENTION

[0003] It can be desirable in the medical and veterinary arts to reducefat accumulation in fat-storing cells (“adipocytes”). In obese human andnon-human animals in particular, adipocytes that accumulate excesslipids can become insulin-resistant, a characteristic that has manyadverse effects including the development of diabetes. Adipocytes cannotdirectly take up and accumulate triglycerides from the blood. Instead,an cell-associated extracellular lipoprotein lipase (LPL) enzyme breaksdown blood-borne triglyceride molecules into a glycerol molecule andthree fatty acids molecules. The adipocytes can take up and convert thefatty acids into triacylglycerides for accumulation and storage.

[0004] Likewise, adipocytes cannot directly secrete fatty acids.Instead, a intracellular hormone-sensitive lipase breaks down cellulartriacylglycerides into free fatty acid and glycerol metabolites that canbe released from the cells. Hormone-sensitive lipase activity increasesand decreases in response to its phosphorylation by a cyclicAMP-dependent protein kinase. The extent of phosphorylation, in turn, isdetermined by cyclic AMP levels. Norepinephrine and glucagon stimulatecyclic AMP production while insulin decreases cyclic AMP levels.

[0005] Conjugated linoleic acid (CLA), a series of positional andgeometric isomers having eighteen carbons and a pair of conjugateddouble bonds, has many biological activities including those related tothe metabolism of body fat and arachidonic acid. CLA inhibits LPLactivity and prevents fat accumulation in human and non-human animals.Other compounds and methods for controlling body fat by reducingaccumulation of fat in fat cells are desired.

[0006] CLA also inhibits platelet aggregation, in that it inhibitscyclooxygenase (COX)-directed conversion of arachidonic acid toprostaglandins and thromboxanes. Thromboxane A2 is a potent inducer ofplatelet aggregation. Inhibition of platelet aggregation advantageouslyreduces the risk of arteriosclerosis, inflammation, and risk of stroke.Millions of Americans are prescribed aspirin to reduce the risk of thesediseases. Other compounds and methods for inhibiting plateletaggregation are also desired.

BRIEF SUMMARY OF THE INVENTION

[0007] The present invention provides a method for inhibitingadipocyte-associated lipoprotein lipase activity in a human or non-humananimal by administering a treating agent that comprises at least one ofconjugated nonadecadienoic acid (CNA) and at least one derivativethereof in an amount effective to modulate the lipoprotein lipaseactivity. As a result, body fat is controlled, cyclooxygenase activityis inhibited or platelet aggregation is inhibited in the animal, therebypreventing or treating a related condition or disease.

[0008] The present invention also provides an edible compositioncomprising at least one of CNA and at least one derivative thereof in anamount effective to modulate the lipoprotein lipase activity, with anedible carrier.

[0009] The present invention also provides a pharmaceutical compositioncomprising at least one of CNA and at least one derivative thereof in anamount effective to modulate the lipoprotein lipase activity, with apharmaceutical carrier.

[0010] It is an object of the present invention to provide compositionsand methods for controlling body fat, inhibiting cyclooxygenaseactivity, reducing release of PGE2, and inhibiting platelet aggregationin a cell of a human or non-human animal.

[0011] Other objects, features, and advantages of the invention will beapparent upon consideration of the following detailed description takenin conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0012]FIG. 1 shows effects of CNA and CLA on LPL activity (A), glycerolrelease (B), cellular triacylglycerol (C), and cellular glycerol (D) in3T3-L1 adipocytes.

[0013]FIG. 2 shows effects of CNA on collagen-induced whole bloodplatelet aggregation of guinea pigs (mean±standard deviation, n=5).

[0014]FIG. 3 shows effects of CNA on ADP-induced whole blood plateletaggregation of guinea pigs (mean±standard deviation, n=5).

[0015]FIG. 4 shows effects of CNA on thromboxane B2 release in wholeblood of guinea pigs stimulated by collagen (mean±standard deviation,n=5).

[0016]FIG. 5 shows effects of CNA on ovine cyclooxygenase-1 activity(mean±standard deviation, n=3).

[0017]FIG. 6 shows effects of CNA on ovine cyclooxygenase-2 activity(mean±standard deviation, n=5).

[0018]FIG. 7 depicts the inhibitory effects of CNA onlipopolysaccharide-stimulated macrophages.

DETAILED DESCRIPTION OF THE INVENTION

[0019] The applicants here disclose biological activities of CNA relatedto the metabolism of body fat and arachidonic acid. In particular, CNAmodulates adipocyte-associated LPL activity and controls body fat levelsin human and non-human animals when administered in sufficient quantity.Further, CNA inhibits cyclooxygenase 1 (COX 1) activity andcyclooxygenase 2 (COX 2) activity as well as thromboxane B2 formationand platelet aggregation in the blood of human and non-human animalswhen administered in sufficient quantity.

[0020] CNA is a 19 carbon free fatty acid having a pair of conjugateddouble bonds. CNA can be prepared by alkali isomerization of c10, c13nonadecadienoic acid (NA, commercially available from Nu-Chek Prep,Elysian, Minn.), using the method described for preparing CLA by Y. Parket al., Lipids 34 (1999) 235-241, incorporated herein by reference as ifset forth in its entirety. CNA can also be prepared using the methods ofJanssen, G., et al., “Location of Double Bonds in Polyenic Long-ChainCarboxylic Acids Containing a Conjugated Diene Unit,” Biomedical andEnvironmental Mas Spectrometry 15:1-6 (1988) and Verhulst, A., et al.,“Isolation of Polyunsaturated Long Chain Fatty Acids byPropionibacteria,” Systematic and Applied Microbiology 9:12-15 (1987),each incorporated herein by reference as if set forth in its entirety.CNA isomers of interest for use in the method include t11, c13 CNA andc10, t12 CNA.

[0021] The treating agent comprises at least one of CNA and at least oneCNA derivative. A CNA derivative is preferred over CNA itself as acomponent of edible and pharmaceutical compositions. A CNA derivative ispreferably an ester, phospholipid or salt of CNA. Preferred esters are aglyceride, a methyl ester and an ethyl ester. Still more preferably, thepreferred glyceride is a triglyceride, diglyceride, or monoglyceride ofCNA. Methods for preparing triglycerides, diglycerides, monoglycerides,phospholipids and salts from free fatty acids are known in the art. Amethod of making triglycerides from fatty acids can be found in Journalof General Chemistry, USSR, 34: 1918, 1964, incorporated by referenceherein as if set forth in its entirety. Briefly in this method, methylesters of fatty acids are mixed with triacetin and sodium methoxide. Themixture is heated in an oil bath at 80° C. for five hours and fatty acidtriglycerides are obtained. Alternatively, free CNA can be fed to ananimal. The animal esterifies CNA and the esters (phospholipids,triglycerides, diglycerides and monoglycerides) can be extracted fromthe blood and separated using methods known in the art. Still anothermethod for making CNA in triglyceride form is the method used inconnection with CLA in U.S. Pat. No. 6,177,580, incorporated herein byreference as if set forth in its entirety. CNA metabolites arestructurally distinct from those of CLA.

[0022] Examples of methods for preparing phospholipids from fatty acidsare found in the following references, each of which is incorporatedherein by reference as if set forth in its entirety: Babaev, et al.,Mater. Vegs. Simp. (1981) Meeting Date 1980, 62-9; Bilimoria, et al, J.Chem. Soc. (1968) (12) 1404-12; Bonsen, et al., Chem. Phys. Lipids(1967) 1(2) 100-9; Isaacson, et al., Chem. Phys. Lipids (1990) 52(3-4)217-26; Smith, et al., Tech. Life Sci., [Sect.]: Biochem. (1982) 8406;and Zhu et al., Journal of Chemical Research, Synopses (1999)(8)500-501.

[0023] CNA salts can be made using the method for making CLA saltsdescribed in U.S. Pat. No. 5,070,104, incorporated herein by referenceas if set forth in its entirety, substituting CNA or derivative thereoffor the CLA or derivative thereof in the patent. Lithium salts areparticularly preferred.

[0024] In one embodiment, the present invention is a method ofinhibiting activity of LPL associated with a cell. “LPL associated witha cell” is LPL located not inside but outside of the adipocyte cell. Themethod involves treating the cell with CNA, a CNA derivative, or acombination of either of the foregoing in an amount effective to inhibitthe associated lipoprotein lipase activity. When treating an in vitrocultured cell, the treating agent can be added into the culture mediumto a total concentration of CNA and/or derivatives thereof in the rangeof from about 10 μM to about 1,000 μM, preferably from about 30 μM toabout 500 μM, and most preferably from about 50 μM to about 150 μM.

[0025] When treating a cell in a human or non-human animal, an amount ofthe treating agent effective to achieve an object of the invention isadministered into the animal by, e.g., feeding, intravenous injection,topical treatment on skin, or direct injection into adipose tissue. Thetreatment can be applied to organs, as well—particularly organsharvested for transplant and having elevated cyclooxygenase enzyme. Forconvenience, feeding is a preferred administration route. When fed, thetotal amount of CNA and/or derivatives thereof in the treating agent canrange from about 0.0001% to about 5% by weight in diet, preferably fromabout 0.05% to about 1% by weight in diet, and most preferably fromabout 0.1% to about 0.5% by weight in diet. One of ordinary skill in theart can readily measure the total concentration of these agents in theblood when different diets listed above are fed and determine thecorresponding dose of these agents for intravenous injection. Whenadministered directly into the adipose tissue, the concentration ofthese agents in the injection solution are the same as the concentrationin the culture medium described above.

[0026] In another embodiment, the present invention is a method ofcontrolling body fat in a human or non-human animal. Body fat controlmeans is achieved when at least one of the following three effects isobserved: the body fat level is reduced in the animal to a level lowerthan that just before CNA treatment; the body fat level is maintained inthe animal at a level substantially the same as that just before CNAtreatment wherein the body fat level was increasing before CNAtreatment; and body fat level remains lower in the animal than it wouldhave been without CNA treatment. The method involves administering CNA,a CNA derivative or a combination thereof into the animal. Theadministration routes and doses are as described above for inhibitingLPL activity.

[0027] In still another embodiment, the present invention is method oftreating a cell to inhibit cyclooxygenase activity in the cell. Bycyclooxygenase activity, we mean the activity of cyclooxygenase-1,cyclooxygenase-2, or both. The inhibition can arise by various means,including but not limited to altering the cyclooxygenase primary,secondary, tertiary or quaternary structure, by reducing or eliminatingthe cyclooxygenase, or by modifying transcription or translation ofcyclooxygenase-encoding polynucleotides. The method involves treatingthe cell with CNA, a CNA derivative or a combination thereof. Whentreating an in vitro cultured cell, tissue or organ, the treating agentcan be added into the culture medium to a total concentration of CNAand/or derivatives thereof in the range of from about 3 μM to about1,000 μM, preferably from about 5 μM to about 500 μM, and mostpreferably from about 10 μM to about 300 μM.

[0028] When treating a cell of a human or non-human animal, an amount ofthe treating agent effective to achieve an object of the invention isadministered into the animal by, e.g., feeding, intravenous injection,topical treatment of skin, or direct injection into adipose tissue. Forconvenience, feeding and topical administration are preferredadministration routes when treating a cell in a human or non-humananimal. Whether fed or injected intravenously, the total concentrationof CNA and/or derivatives in the treating agent is adjusted such thatafter injection, the total concentration of CNA and/or derivatives inthe blood ranges from 3 μM to about 1,000 μM, preferably from about 5 μMto about 500 μM, and most preferably from about 10 μM to about 300 μM.Given the stated blood concentration ranges, one of ordinary skill inthe art can readily determine the dose to be administered.

[0029] In yet another embodiment, the present invention is a method ofinhibiting platelet aggregation, or preventing or treating a conditionor disease related to platelet aggregation in a human or non-humananimal. Examples of conditions or diseases related to plateletaggregation include but are not limited to arteriosclerosis,inflammation and stroke. The method involves administering CNA, a CNAderivative or a combination thereof into the animal. Suitableadministration routes and doses are as described above for inhibitingcyclooxygenase activity.

[0030] In another embodiment, the present invention is an ediblecomposition suitable for consumption by a human or non-human animal,particularly for use in the disclosed methods. The composition containsCNA or a CNA derivative with an edible carrier of the type known to theart. The presence of the carrier should not unduly interfere with theactivity of the CNA or CNA derivative in the methods of the invention.Thus, the carrier should not reduce the effect of the CNA or CNAderivative in a method by more than 20 percent. It is understood that ifthe carrier masks the activity in the methods, the amount of CNA or CNAderivative can be increased accordingly. It is understood that theedible compositions of the invention include foods having added theretoat least one of CNA and at least one CNA derivative. Particularlyadvantageous foods include food products that contain fatty acids suchas dairy products and nutritional supplements.

[0031] In another embodiment, the present invention is a pharmaceuticalcomposition suitable for administration to a human or non-human animal,particularly for use in the disclosed methods. The composition containsat least one of CNA or a CNA derivative with an pharmaceutical carrierof the type known to the art. The presence of the carrier should notunduly interfere with the activity of the CNA or CNA derivative in themethods of the invention. Thus, the carrier should not reduce the effectof the CNA or CNA derivative in a method by more than 20 percent. It isunderstood that if the carrier masks the activity in the methods, theamount of CNA or CNA derivative can be increased accordingly.

[0032] One of ordinary skill in the art is familiar with edible andpharmaceutical formulations that are suitable for human and non-humananimal use.

[0033] The invention will be more fully understood upon consideration ofthe following non-limiting examples.

EXAMPLE 1

[0034] Effects of CNA and CLA on heparin-releasable LPL activity,glycerol release, cellular triacylglycerol, and cellular glycerol in3T3-L1 adipocytes

[0035] Materials and Methods

[0036] CNA was synthesized from NA (purchased from Nu-Chek Corp.,Elysian, Minn.), using the method described in Park, Y. et al., Lipids34 (1999) 235-241 for producing CLA from linoleic acid; the CNA productsynthesized contained 97.3% CNA (c10, t12, 33.4%; t11, c13, 32.9%; t10,t12/t11, t13, 23.4%; other isomers, 7.6%) with the remainder asunreacted NA. CLA was synthesized as described by Park, et al., supra;the CLA product synthesized contained 97.8% CLA (c9, t11/t9, c11, 45.7%;t10, c12, 47.6%; t9, t11/t10, t12, 1.7%; others, 2.8%) with theremainder as unreacted linoleic acid.

[0037] Cultured cells were treated with fatty acid-albumin complexes for2 days, beginning at day 6 post-differentiation. Data were analyzed aslog value with a two-way ANOVA using fatty acid (as treatments) andexperiment. If the interaction between treatment and experiment wassignificant, this interaction was used as the error term. Additionalexperimental details were as described in Park, et al., supra.

[0038] Results

[0039] Treatment values indicate the concentration of test materials(μM); all treatment groups included albumin at 100 μM. Reported valuesare mean±S.E. (n=8, collected from two independent experiments). Meanswith different letters are significantly different (P<0.05). CNAinhibited LPL activity in cultured 3T3-L1 adipocytes (FIG. 1A). Inaddition, CNA enhanced glycerol release (FIG. 1B) and reducedintracellular triacylglycerol (FIG. 1C) and glycerol (FIG. 1D) in thesame cells. In contrast, NA at 50 or 100 μM was ineffective.

EXAMPLE 2 Effects of CNA on Body Composition

[0040] Materials and Methods

[0041] Female ICR mice (4-week-old) and semi-purified diets (TD94060,99% basal mix) were purchased from Harlan Sprague Dawley (Madison, Wis.)and Harlan Teklad (Madison, Wis.), respectively. CLA was obtained fromNatural Lipids Ltd AS, Hovdebygda, Norway; its composition was 88.7% CLA(c9, t11/t9, c11, 41.9%; t10, c12, 43.5%; t9, t11/10, t12, 1.5%; others,1.8%) with the remainder as oleic acid, 5.6%; palmitic acid, 1.4%; andlinoleic acid, 0.5%. CNA was kindly provided by the US Food and DrugAdministration; it was 98.6% CNA (c10, t12, 42.3%; t11, c13, 48.0%; t10,t12/t11, t13, 5.1%; others, 3.2%) with the remainder as unreacted NA.CLA or CNA was added to diets at the expense of corn oil to maintain5.5% fat. Diet was stored at −20° C. until use. Mice were housedindividually in a windowless room with a 12-h light-dark cycle in strictaccordance to guidelines established by the Research Animal ResourcesCenter of University of Wisconsin-Madison. Diet and water, available adlibitum, were freshly provided three times per week.

[0042] After a 5 day adaptation period, mice were randomly separatedinto 3 groups and fed control diet, diet supplemented with 0.3% CLA, ordiet supplemented with 0.3% CNA for 2 weeks. 0.3% of diet CLA is asub-optimal dose for CLA's effects on body composition.

[0043] Results

[0044] Results are mean±S.E. (n=6). Means with different letters in eachcolumn are significantly different (P<0.01 for water and fat, and P<0.05for empty carcass weight (ECW), protein and ash).

[0045] Table 1 shows that a CNA diet in mice reduced body fat mass gainand enhanced the amount of whole body water, protein and ash. Feedingmice with CLA diet caused similar but less dramatic changes.

[0046] The CLA and CNA groups also displayed reductions in body weightgain and feed intake relative to controls during the experimentalperiod. Body weight gain per animal for controls, CLA and CNA groupswere 2.5 g, 1.3 g, 0.2 g, respectively; total feed intake per group forcontrols, CLA and CNA groups were 57.9 g, 51.0 g, 47.1 g, respectively.It has been repeatedly shown in CLA-feeding trials (see, e.g., Y. Park,et al., Lipids 34 (1999) 243-248) that these are transient effects thatare sometimes but not always seen, and are unrelated to the observedchanges in body composition. TABLE 1 Effects of CNA and CLA on bodycomposition. ECW (g) % Fat % Water % Protein % Ash Control 24.5^(a) ±0.7 20.00^(a) ± 2.28 57.9^(a) ± 1.7 18.35^(a) ± 0.61 3.83^(a) ± 0.15 CLA23.2^(ab) ± 0.6  14.98^(a) ± 2.08 59.9^(a) ± 1.7 19.16^(a) ± 0.504.01^(ab) ± 0.13  CAN 21.8^(b) ± 0.4  3.90^(b) ± 0.44 66.2^(b) ± 0.420.79^(b) ± 0.24 4.42^(b) ± 0.12

EXAMPLE 3 Effects of CNA on Platelet Aggregation, Thromboxane B2 Level,PGE2 Level, and COX-1 and COX-2 Activity

[0047] Materials and Methods

[0048] Female guinea pigs and diets were purchased from Harlan Teklad(Madison, Wis). Whole Blood Aggregometer, collagen and ADP werepurchased from Chrono-log Corp. (Havertown, Pa.). All ELISA kits werepurchased from Cayman Chemical (Ann Arbor, Mich.).

[0049] Whole blood platelet aggregation: Guinea Pigs were anesthetizedwith a combination of ketamine (35 mg/kg) and xylazine (10 mg/kg) bysubcutaneous administration. Heparinized blood (20 unit/ml) was obtainedby cardiac puncture. Within 5-10 minutes, the blood (1.0 ml) waspreheated at 37° C. and then incubated with free fatty acid or DMSO(control) for 2 minutes. Collagen (4 μg/ml) or ADP (10 μM) was nextadded to the blood to start the platelet aggregation process. Plateletaggregation was measured using an Aggregometer.

[0050] Determination of thromboxane B2 level by ELISA: After the wholeblood platelet aggregation, the collagen-stimulated blood wascentrifuged for plasma and stored at −80° C. The plasma thromboxanelevel was determined by ELISA according to the manufacturer'sinstructions. Inhibitory effects on ovine COX-1 and COX-2: COX-1 orCOX-2, heme and reaction buffer were pre-incubated under 37° C. CNA orDMSO (control) was added to the system and co-incubated for 5 minutes.Arachidonic acid was added and the reaction was maintained at 37° C. for2 minutes. The catalysis reaction was terminated by adding HCL. Totalprostanoids were measured by ELISA according to manufacturer'sinstructions.

[0051] Results

[0052] CNA inhibited the collagen- and ADP-stimulated plateletaggregation in a dose-dependent manner (FIG. 2 and FIG. 3). CNA alsoreduced thromboxane B2 level in collagen-stimulated blood in adose-dependent manner (FIG. 4). In addition, CNA inhibited pure ovineCOX-1 and COX-2 in a dose-dependent manner (FIG. 5 and FIG. 6).

[0053] As a further indication of the ability of CNA to inhibit COXenzymes, FIG. 7 depicts the inhibition of PGE2 released from stimulatedmacrophages in the presence of varying amounts of CNA. Confluentmacrophages were incubated with CNA or linoleic acid (LA; control) atthe concentrations shown for 24 hours in 0.5% fetal bovine serum thenwere stimulated with 100 ng/ml lipopolysaccharide (LPS) for 8 hours. Thelevel of PGE2 in the medium was determined by ELISA assay. FIG. 7 showssubstantial inhibition to less than 1000 pg/ml from a level of about3000 pg/ml in LPS-stimulated macrophages without CNA.

[0054] The present invention is not intended to be limited to theforegoing examples, but encompasses all such modifications andvariations as come within the scope of the appended claims.

We claim:
 1. A method for modulating lipoprotein lipase activityassociated with a cell, the method comprising the step of: administeringto the cell a composition that comprises at least one treating agentselected from the group consisting of a conjugated nonadecadienoic acid(CNA) and at least one CNA derivative in an amount sufficient tomodulate the lipoprotein lipase activity.
 2. The method of claim 1wherein the cell is an in vitro cultured cell.
 3. The method of claim 1wherein the cell is in a human or non-human animal.
 4. The method ofclaim 1 wherein the at least one CNA derivative is selected from thegroup consisting of an ester of CNA, a phospholipid of CNA and a salt ofCNA.
 5. The method of claim 4 wherein the ester of CNA is a glyceride.6. The method of claim 5 wherein the glyceride is selected from thegroup consisting of a triglyceride of CNA, a diglyceride of CNA, and amonoglyceride of CNA.
 7. A method for controlling body fat in a human ornon-human animal, the method comprising the step of: administering acomposition that comprises at least one treating agent selected from thegroup consisting of a conjugated nonadecadienoic acid (CNA) and at leastone CNA derivative in an amount sufficient to control body fat in theanimal.
 8. The method of claim 7 wherein the administering stepcomprises oral administration.
 9. The method of claim 7 wherein theadministering step comprises intravenous injection.
 10. The method ofclaim 7 wherein the administering step comprises direct injection intoadipose tissue.
 11. The method of claim 7 wherein the administering stepcomprises topical treatment of skin.
 12. The method of claim 7 whereinthe at least one CNA derivative is selected from the group consisting ofan ester of CNA, a phospholipid of CNA and a salt of CNA.
 13. The methodof claim 12 wherein the ester of CNA is a glyceride.
 14. The method ofclaim 13 wherein the glyceride is selected from the group consisting ofa triglyceride of CNA, a diglyceride of CNA, and a monoglyceride of CNA.15. A method for inhibiting cyclooxygenase activity in a cell, themethod comprising the step of: administering to the cell a compositionthat comprises at least one treating agent selected from the groupconsisting of a conjugated nonadecadienoic acid (CNA) and at least oneCNA derivative in an amount sufficient to inhibit cyclooxygenaseactivity.
 16. The method of claim 15 wherein the cell is an in vitrocultured cell.
 17. The method of claim 15 wherein the cell is in a humanor non-human animal.
 18. The method of claim 15 wherein the at least oneCNA derivative is selected from the group consisting of an ester of CNA,a phospholipid of CNA and a salt of CNA.
 19. The method of claim 18wherein the ester of CNA is a glyceride.
 20. The method of claim 19wherein the glyceride is selected from the group consisting of atriglyceride of CNA, a diglyceride of CNA, and a monoglyceride of CNA.21. A method for inhibiting platelet aggregation in a human or non-humananimal, the method comprising the step of: administering to the cell acomposition that comprises at least one treating agent selected from thegroup consisting of a conjugated nonadecadienoic acid (CNA) and at leastone CNA derivative in an amount sufficient to inhibit plateletaggregation.
 22. The method of claim 21 wherein the administering stepcomprises oral administration.
 23. The method of claim 21 wherein theadministering step comprises intravenous injection.
 24. The method ofclaim 21 wherein the administering step comprises topical treatment ofskin.
 25. The method of claim 21 wherein the at least one CNA derivativeis selected from the group consisting of an ester of CNA, a phospholipidof CNA and a salt of CNA.
 26. The method of claim 25 wherein the esterof CNA is a glyceride.
 27. The method of claim 26 wherein the glycerideis selected from the group consisting of a triglyceride of CNA, adiglyceride of CNA, and a monoglyceride of CNA.
 28. A method forpreventing or treating a condition or disease related to plateletaggregation in a human or non-human animal, the method comprising thestep of: administering to the cell a composition that comprises at leastone treating agent selected from the group consisting of a conjugatednonadecadienoic acid (CNA) and at least one CNA derivative in an amountsufficient to prevent or treat the condition or disease.
 29. The methodof claim 28 wherein the administering step comprises oraladministration.
 30. The method of claim 28 wherein the administeringstep comprises intravenous injection.
 31. The method of claim 28 whereinthe administering step comprises topical treatment of skin.
 32. Themethod of claim 28 wherein the at least one CNA derivative is selectedfrom the group consisting of an ester of CNA, a phospholipid of CNA anda salt of CNA.
 33. The method of claim 32 wherein the ester of CNA is aglyceride.
 34. The method of claim 33 wherein the glyceride is selectedfrom the group consisting of a triglyceride of CNA, a diglyceride ofCNA, and a monoglyceride of CNA.
 35. A composition comprising: at leastone treating agent selected from the group consisting of a conjugatednonadecadienoic acid (CNA) and at least one CNA derivative; and anedible carrier.
 36. The composition of claim 35 wherein the at least oneCNA derivative is selected from the group consisting of an ester of CNA,a phospholipid of CNA and a salt of CNA.
 37. The composition of claim 36wherein the ester of CNA is a glyceride.
 38. The composition of claim 37wherein the glyceride is selected from the group consisting of atriglyceride of CNA, a diglyceride of CNA, and a monoglyceride of CNA.39. A composition comprising: at least one treating agent selected fromthe group consisting of a conjugated nonadecadienoic acid (CNA) and atleast one CNA derivative; and a pharmaceutical carrier.
 40. Thecomposition of claim 39 wherein the at least one CNA derivative isselected from the group consisting of an ester of CNA, a phospholipid ofCNA and a salt of CNA.
 41. The composition of claim 40 wherein the esterof CNA is a glyceride.
 42. The composition of claim 41 wherein theglyceride is selected from the group consisting of a triglyceride ofCNA, a diglyceride of CNA, and a monoglyceride of CNA.